Prodrugs of a2b adenosine receptor antagonists

ABSTRACT

Disclosed are prodrugs of A 2B  adenosine receptor antagonists, and their use in treating mammals for various disease states.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a Continuation of U.S. patent application Ser. No.11/453,414, filed Jun. 14, 2006, which claims priority to U.S.Provisional Patent Application Ser. No. 60/691,408, filed Jun. 16, 2005,the complete disclosure of which is hereby incorporated by reference.

FIELD OF THE INVENTION

The present invention relates to prodrugs of A_(2B) adenosine receptorantagonists, and to their use in treating mammals for various diseasestates, such as gastrointestinal disorders, immunological disorders,hypersensitivity disorders, neurological disorders, and cardiovasculardiseases due to both cellular hyperproliferation and apoptosis, and thelike. The invention also relates to methods for the preparation of suchcompounds, and to pharmaceutical compositions containing them.

BACKGROUND

Adenosine is a naturally occurring nucleoside, which exerts itsbiological effects by interacting with a family of adenosine receptorsknown as A₁, A_(2A), A_(2B), and A₃, all of which modulate importantphysiological processes. For example, A_(2A) adenosine receptorsmodulate coronary vasodilation, A_(2B) receptors have been implicated inmast cell activation, asthma, vasodilation, regulation of cell growth,intestinal function, and modulation of neurosecretion (See AdenosineA_(2B) Receptors as Therapeutic Targets, Drug Dev Res 45:198; Feoktistovet al., Trends Pharmacol Sci 19:148-153), and A₃ adenosine receptorsmodulate cell proliferation processes.

Adenosine A_(2B) receptors are ubiquitous, and regulate multiplebiological activities. For example, adenosine binds to A_(2B) receptorson endothelial cells, thereby stimulating angiogenesis. Adenosine alsoregulates the growth of smooth muscle cell populations in blood vessels.Adenosine stimulates A_(2B) receptors on mast cells, thus modulatingType I hypersensitivity reactions. Adenosine also stimulatesgastrosecretory activity by ligation with A_(2B) in the intestine.

While many of these biological effects of adenosine are necessary tomaintain normal tissue homeostasis, under certain physiological changesit is desirable to modulate its effects. For example, the binding ofA_(2B) receptors stimulates angiogenesis by promoting the growth ofendothelial cells. Such activity is necessary in healing wounds, but thehyperproliferation of endothelial cells promotes diabetic retinopathy.Also, an undesirable increase in blood vessels occurs in neoplasia.Accordingly, inhibition of the binding of adenosine to A_(2B) receptorsin the endothelium will alleviate or prevent hypervasculation, thuspreventing retinopathy and inhibiting tumor formation.

A_(2B) receptors are found in the colon in the basolateral domains ofintestinal epithelial cells, and when acted upon by the appropriateligand act to increase chloride secretion, thus causing diarrhea, whichis a common and potentially fatal complication of infectious diseasessuch as cholera and typhus. A_(2B) antagonists can therefore be used toblock intestinal chloride secretion, and are thus useful in thetreatment of inflammatory gastrointestinal tract disorders, includingdiarrhea.

Insensitivity to insulin exacerbates diabetes and obesity. Insulinsensivity is decreased by the interaction of adenosine with A_(2B)receptors. Thus, blocking the adenosine A_(2B) receptors of individualswith diabetes or obesity would benefit patients with these disorders.

Another adverse biological effect of adenosine acting at the A_(2B)receptor is the over-stimulation of cerebral IL-6, a cytokine associatedwith dementias and Alzheimer's disease. Inhibiting the binding ofadenosine to A_(2B) receptors would therefore mitigate thoseneurological disorders that are produced by IL-6.

Type I hypersensitivity disorders, such as chronic obstructive pulmonarydisease (COPD), asthma, hay fever, and atopic eczema, are stimulated bymast cells binding to A_(2B)-receptors. Accordingly, blocking suchadenosine receptors provides a therapeutic benefit against suchdisorders.

There are several compounds presently used in the treatment of asthma.For example, theophylline is an effective anti-asthmatic agent, eventhough it is a poor adenosine receptor antagonist. However, high plasmalevels are needed for it to be effective. Additionally, theophylline hassubstantial side effects, most of which are due to its CNS action, whichprovide no beneficial effects in the treatment of asthma, and to thefact that it non-specifically blocks all adenosine receptor subtypes.

Additionally adenosine treatment, such as inhaled adenosine (oradenosine monophosphate), provokes bronchoconstriction in asthmatics,but not in the normal population. This process is known to involve mastcell activation, in that it releases mast cell mediators, includinghistamine, PGD2-β-hexosaminidase and tryptase. This response is blockedby specific histamine H₁ blockers and chromolyn sodium. Accordingly,there is an intrinsic difference in the way adenosine interacts withmast cells from asthmatics, and thus A_(2B) antagonists are particularlyuseful in modulating mast cell function or in the activation of humanlung cells.

U.S. Pat. No. 6,825,349 discloses novel A_(2B) adenosine receptorantagonists that are potent and selective for the A_(2B) adenosinereceptor. A category of preferred compounds disclosed in the abovepatent application has been identified in which the 7-position of thexanthine moiety is unsubstituted. Such compounds are known to berelatively insoluble in aqueous media and difficult to formulate usingconventional pharmaceutical excipients, and thus potentially difficultto formulate in a manner that provides reproducible plasma levels of thecompound undergoing evaluation in mammals, in particular humans. We havediscovered compounds that are more soluble in aqueous media and/orconventional pharmaceutical excipients, and are surprisingly active asprodrugs of the compounds of '349.

SUMMARY OF THE INVENTION

U.S. Pat. No. 6,825,349 discloses novel A_(2B) adenosine receptorantagonists. One embodiment of the invention of '349 is represented bythe following formula:

in which:

-   R¹ and R² are independently lower alkyl;-   R³ is hydrogen or optionally substituted alkyl; and-   R⁴ is optionally substituted phenyl;-   and the pharmaceutically acceptable salts thereof.

One preferred embodiment of compounds within the scope of Formula Aincludes those compounds in which the 7-position of the xanthine moietyis unsubstituted; that is, where R³ is hydrogen. Particularly preferredare those compounds in which R^(I) and R² are different, and are loweralkyl, and R⁴ is 3-trifluoromethylphenyl. However, it has been foundthat the preferred compounds are relatively insoluble in aqueous mediaand difficult to formulate using conventional pharmaceutical excipients,and thus potentially difficult to formulate in a manner that providesreproducible plasma levels of a compound undergoing evaluation inmammals, in particular humans. It has surprisingly been discovered thata small subset of compounds of Formula A behave as prodrugs of thepreferred compounds. These compounds are chosen from compounds ofFormula A in which R³ is substituted methyl; in particular, thosecompounds in which the substitution on the methyl provides an ester or aphosphate derivative. Such compounds are more soluble in aqueous mediaand/or conventional pharmaceutical excipients than the compounds ofFormula A, and provide higher plasma levels of the active moiety (thosecompounds of Formula A in which R³ is hydrogen) than administration ofthe active moiety itself.

Accordingly, in a first aspect, the present invention relates toprodrugs of Formula I having the formula:

wherein:

-   R¹ and R² are independently lower alkyl;-   R⁴ is optionally substituted phenyl;-   X is hydrogen or methyl; and-   Y is —C(O)R, in which R is independently optionally substituted    lower alkyl, optionally substituted aryl, or optionally substituted    heteroaryl; or-   Y is —P(O)(OR⁵)₂, in which R⁵ is hydrogen or lower alkyl optionally    substituted by phenyl or heteroaryl;-   and the pharmaceutically acceptable salts thereof.

One preferred group of compounds of Formula I are those in which R¹ andR² are ethyl or n-propyl, especially those compounds in which R¹ isn-propyl and R² is ethyl. Preferably R⁴ is 3-(trifluoromethyl)phenyl andX is hydrogen.

One preferred subgroup includes those compounds of Formula I in which Yis —C(O)R, particularly those compounds in which R is methyl, ethyl,n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, or n-pentyl, moreparticularly where R is methyl, n-propyl, or t-butyl. Another preferredsubgroup includes those compounds of Formula I in which Y is —P(O)OR⁵)₂, especially where R⁵ is hydrogen.

In a second aspect, the present invention relates to a method of usingthe compounds of Formula I for treating a disease state in a mammal thatis alleviable by treatment with an A_(2B) adenosine receptor antagonist,in particular atherosclerosis, angiogenesis, diabetic retinopathy,cancer, chronic obstructive pulmonary disease, or asthma, orinflammatory gastrointestinal tract disorders such as diarrhea, orneurological disorder such as senile dementia, Alzheimer's disease, orParkinson's disease.

A third aspect of this invention relates to methods for preparing thecompounds of Formula I.

A fourth aspect of this invention relates to pharmaceuticalformulations, comprising a therapeutically effective amount of acompound of Formula I and at least one pharmaceutically acceptableexcipient.

At present, the preferred compounds are:

[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methylacetate;

[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl2,2-dimethylpropanoate;

[3-ethyl-2,6-dioxo-l-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methylbutanoate; and

[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}-pyrazol-4-yl)(1,3,7-trihydropurin-7-yl]methyldihydrogen phosphate.

Definitions and General Parameters

As used in the present specification, the following words and phrasesare generally intended to have the meanings as set forth below, exceptto the extent that the context in which they are used indicatesotherwise.

The term “alkyl” refers to a monoradical branched or unbranchedsaturated hydrocarbon chain having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19 or 20 carbon atoms. This term isexemplified by groups such as methyl, ethyl, n-propyl, iso-propyl,n-butyl, iso-butyl, t-butyl, n-hexyl, n-decyl, tetradecyl, and the like.

The term “substituted alkyl” refers to:

-   -   1) an alkyl group as defined above, having 1, 2, 3, 4 or 5        substituents, preferably 1 to 3 substituents, selected from the        group consisting of alkenyl, alkynyl, alkoxy, cycloalkyl,        cycloalkenyl, acyl, acylamino, acyloxy, amino, aminocarbonyl,        alkoxycarbonylamino, azido, cyano, halogen, hydroxy, keto,        phosphate, thiocarbonyl, carboxy, carboxyalkyl, arylthio,        heteroarylthio, heterocyclylthio, thiol, alkylthio, aryl,        aryloxy, heteroaryl, aminosulfonyl, aminocarbonylamino,        heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino,        alkoxyamino, nitro, —SO-alkyl, —SO-aryl, —SO-heteroaryl,        —SO₂-alkyl, SO₂-aryl and —SO₂-heteroaryl. Unless otherwise        constrained by the definition, all substituents may optionally        be further substituted by 1, 2, or 3 substituents chosen from        alkyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy,        halogen, CF₃, amino, substituted amino, cyano, and —S(O)_(n)R,        where R is alkyl, aryl, or heteroaryl and n is 0, 1 or 2; or    -   2) an alkyl group as defined above that is interrupted by 1-10        atoms independently chosen from oxygen, sulfur and NR_(a)—,        where R_(a) is chosen from hydrogen, alkyl, cycloalkyl, alkenyl,        cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclyl. All        substituents may be optionally further substituted by alkyl,        alkoxy, halogen, CF₃, amino, substituted amino, cyano, or        —S(O)_(n)R, in which R is alkyl, aryl, or heteroaryl and n is 0,        1 or 2; or    -   3) an alkyl group as defined above that has both 1, 2, 3, 4 or 5        substituents as defined above and is also interrupted by 1-10        atoms as defined above.

The term “lower alkyl” refers to a monoradical branched or unbranchedsaturated hydrocarbon chain having 1, 2, 3, 4, 5, or 6carbon atoms. Thisterm is exemplified by groups such as methyl, ethyl, n-propyl,iso-propyl, n-butyl, iso-butyl, t-butyl, n-hexyl, and the like.

The term “substituted lower alkyl” refers to lower alkyl as definedabove having 1 to 5 substituents, preferably 1, 2, or 3 substituents, asdefined for substituted alkyl, or a lower alkyl group as defined abovethat is interrupted by 1, 2, 3, 4, or 5 atoms as defined for substitutedalkyl, or a lower alkyl group as defined above that has both 1, 2, 3, 4or 5 substituents as defined above and is also interrupted by 1, 2, 3,4, or 5 atoms as defined above.

The term “alkylene” refers to a diradical of a branched or unbranchedsaturated hydrocarbon chain, having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19 or 20 carbon atoms, preferably 1-10carbon atoms, more preferably 1, 2, 3, 4, 5 or 6 carbon atoms. This termis exemplified by groups such as methylene (—CH₂—), ethylene (—CH₂CH₂—),the propylene isomers (e.g., —CH₂CH₂CH₂— and —CH(CH₃)CH₂—) and the like.

The term “lower alkylene” refers to a diradical of a branched orunbranched saturated hydrocarbon chain, preferably having from 1, 2, 3,4, 5, or 6 carbon atoms.

The term “lower alkylene” refers to a diradical of a branched orunbranched saturated hydrocarbon chain, preferably having from 1, 2, 3,4, 5, or 6 carbon atoms.

The term“substituted alkylene” refers to:

-   -   (1) an alkylene group as defined above having 1, 2, 3, 4, or 5        substituents selected from the group consisting of alkyl,        alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl,        acylamino, acyloxy, amino, aminocarbonyl, alkoxycarbonylamino,        azido, cyano, halogen, hydroxy, keto, thiocarbonyl, carboxy,        carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio, thiol,        alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl,        aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy,        hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-aryl,        —SO-heteroaryl, —SO₂-alkyl, SO₂-aryl and —SO₂-heteroaryl. Unless        otherwise constrained by the definition, all substituents may        optionally be further substituted by 1, 2, or 3 substituents        chosen from alkyl, carboxy, carboxyalkyl, aminocarbonyl,        hydroxy, alkoxy, halogen, CF₃, amino, substituted amino, cyano,        and —S(O)_(n)R, where R is alkyl, aryl, or heteroaryl and n is        0, 1 or 2; or    -   (2) an alkylene group as defined above that is interrupted by        1-20atoms independently chosen from oxygen, sulfur and NR_(a)—,        where R_(a) is chosen from hydrogen, optionally substituted        alkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl and        heterocycyl, or groups selected from carbonyl, carboxyester,        carboxyamide and sulfonyl; or    -   (3) an alkylene group as defined above that has both 1, 2, 3, 4        or 5 substituents as defined above and is also interrupted by        1-20 atoms as defined above. Examples of substituted alkylenes        are chloromethylene (—CH(Cl)—), aminoethylene (—CH(NH₂)CH₂—),        methylaminoethylene (—CH(NHMe)CH₂—), 2-carboxypropylene isomers        (—CH₂CH(CO₂H)CH₂—), ethoxyethyl (—CH₂CH₂O—CH₂CH₂—),        ethylmethylaminoethyl        (—CH₂CH₂N(CH₃)CH₂CH₂—),1-ethoxy-2-(2-ethoxy-ethoxy)ethane        (—CH₂CH₂O—CH₂CH₂—OCH₂CH₂—OCH₂CH₂—), and the like.

The term “aralkyl” refers to an aryl group covalently linked to analkylene group, where aryl and alkylene are defined herein. “Optionallysubstituted aralkyl” refers to an optionally substituted aryl groupcovalently linked to an optionally substituted alkylene group. Sucharalkyl groups are exemplified by benzyl, phenylethyl,3-(4-methoxyphenyl)propyl, and the like.

The term “alkoxy” refers to the group R—O—, where R is optionallysubstituted alkyl or optionally substituted cycloalkyl, or R is a group—Y—Z, in which Y is optionally substituted alkylene and Z is optionallysubstituted alkenyl, optionally substituted alkynyl; or optionallysubstituted cycloalkenyl, where alkyl, alkenyl, alkynyl, cycloalkyl andcycloalkenyl are as defined herein. Preferred alkoxy groups areoptionally substituted alkyl-O— and include, by way of example, methoxy,ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy, sec-butoxy,n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, trifluoromethoxy, and the like.

The term “alkylthio” refers to the group R—S—, where R is as defined foralkoxy.

The term “alkenyl” refers to a monoradical of a branched or unbranchedunsaturated hydrocarbon group preferably having from 2 to 20 carbonatoms, more preferably 2 to 10 carbon atoms and even more preferably 2to 6 carbon atoms and having 1-6, preferably 1, double bond (vinyl).Preferred alkenyl groups include ethenyl or vinyl (—CH═CH₂), 1-propyleneor allyl (—CH₂CH═CH₂), isopropylene (—C(CH₃)═CH₂),bicyclo[2.2.1]heptene, and the like. In the event that alkenyl isattached to nitrogen, the double bond cannot be alpha to the nitrogen.

The term “lower alkenyl” refers to alkenyl as defined above having from2 to 6 carbon atoms.

The term “substituted alkenyl” refers to an alkenyl group as definedabove having 1, 2, 3, 4 or 5 substituents, and preferably 1, 2, or 3substituents, selected from the group consisting of alkyl, alkenyl,alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy,amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano, halogen,hydroxy, keto, thiocarbonyl, carboxy, carboxyalkyl, arylthio,heteroarylthio, heterocyclylthio, thiol, alkylthio, aryl, aryloxy,heteroaryl, aminosulfonyl, aminocarbonylamino, heteroaryloxy,heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro,—SO-alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl, SO₂-aryl and—SO₂-heteroaryl. Unless otherwise constrained by the definition, allsubstituents may optionally be further substituted by 1, 2, or 3substituents chosen from alkyl, carboxy, carboxyalkyl, aminocarbonyl,hydroxy, alkoxy, halogen, CF₃, amino, substituted amino, cyano, and—S(O)_(n)R, where R is alkyl, aryl, or heteroaryl and n is 0, 1 or 2.

The term “alkynyl” refers to a monoradical of an unsaturatedhydrocarbon, preferably having from 2 to 20 carbon atoms, morepreferably 2 to 10 carbon atoms and even more preferably 2 to 6 carbonatoms and having at least 1 and preferably from 1-6 sites of acetylene(triple bond) unsaturation. Preferred alkynyl groups include ethynyl,(—C≡CH), propargyl (or prop-1-yn-3-yl, —CH₂C≡CH), and the like. In theevent that alkynyl is attached to nitrogen, the triple bond cannot bealpha to the nitrogen.

The term “substituted alkynyl” refers to an alkynyl group as definedabove having 1, 2, 3, 4 or 5 substituents, and preferably 1, 2, or 3substituents, selected from the group consisting of alkyl, alkenyl,alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy,amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano, halogen,hydroxy, keto, thiocarbonyl, carboxy, carboxyalkyl, arylthio,heteroarylthio, heterocyclylthio, thiol, alkylthio, aryl, aryloxy,heteroaryl, aminosulfonyl, aminocarbonylamino, heteroaryloxy,heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro,—SO-alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl, SO₂-aryl and—SO₂-heteroaryl. Unless otherwise constrained by the definition, allsubstituents may optionally be further substituted by 1, 2, or 3substituents chosen from alkyl, carboxy, carboxyalkyl, aminocarbonyl,hydroxy, alkoxy, halogen, CF₃, amino, substituted amino, cyano, and—S(O)_(n)R, where R is alkyl, aryl, or heteroaryl and n is 0, 1 or 2.

The term “aminocarbonyl” refers to the group —C(O)NRR where each R isindependently hydrogen, alkyl, aryl, heteroaryl, heterocyclyl or whereboth R groups are joined to form a heterocyclic group (e.g.,morpholino). Unless otherwise constrained by the definition, allsubstituents may optionally be further substituted by 1-3 substituentschosen from alkyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy,alkoxy, halogen, CF₃, amino, substituted amino, cyano, and —S(O)_(n)R,where R is alkyl, aryl, or heteroaryl and n is 0, 1 or 2.

The term “acylamino” refers to the group —NRC(O)R where each R isindependently hydrogen, alkyl, aryl, heteroaryl, or heterocyclyl. Unlessotherwise constrained by the definition, all substituents may optionallybe further substituted by 1-3 substituents chosen from alkyl, carboxy,carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF₃, amino,substituted amino, cyano, and —S(O)_(n)R, where R is alkyl, aryl, orheteroaryl and n is 0, 1 or 2.

The term “aryloxy” refers to the groups —O(O)C-alkyl, —O(O)C-cycloalkyl,—O(O)C-aryl, —O(O)C-heteroaryl, and —O(O)C-heterocyclyl. Unlessotherwise constrained by the definition, all substituents may beoptionally further substituted by alkyl, carboxy, carboxyalkyl,aminocarbonyl, hydroxy, alkoxy, halogen, CF₃, amino, substituted amino,cyano, or —S(O)_(n)R, where R is alkyl, aryl, or heteroaryl and n is 0,1 or 2.

The term “aryl” refers to an aromatic carbocyclic group of 6 to 20carbon atoms having a single ring (e.g., phenyl) or multiple rings(e.g., biphenyl), or multiple condensed (fused) rings (e.g., naphthyl oranthryl). Preferred aryls include phenyl, naphthyl and the like.

The term “arylene” refers to a diradical of an aryl group as definedabove. This term is exemplified by groups such as 1,4-phenylene,1,3-phenylene, 1,2-phenylene, 1,4′-biphenylene, and the like.

Unless otherwise constrained by the definition for the aryl or arylenesubstituent, such aryl or arylene groups can optionally be substitutedwith from 1 to 5 substituents, preferably 1 to 3 substituents, selectedfrom the group consisting of alkyl, alkenyl, alkynyl, alkoxy,cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy, amino,aminocarbonyl, alkoxycarbonylamino, azido, cyano, halogen, hydroxy,keto, thiocarbonyl, carboxy, carboxyalkyl, arylthio, heteroarylthio,heterocyclylthio, thiol, alkylthio, aryl, aryloxy, heteroaryl,aminosulfonyl, aminocarbonylamino, heteroaryloxy, heterocyclyl,heterocyclooxy, hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-aryl,—SO-heteroaryl, —SO₂-alkyl, SO₂-aryl and —SO₂-heteroaryl. Unlessotherwise constrained by the definition, all substituents may optionallybe further substituted by 1-3 substituents chosen from alkyl, carboxy,carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF₃, amino,substituted amino, cyano, and —S(O)_(n)R, where R is alkyl, aryl, orheteroaryl and n is 0, 1 or 2.

The term “aryloxy” refers to the group aryl-O— wherein the aryl group isas defined above, and includes optionally substituted aryl groups asalso defined above. The term “arylthio” refers to the group R—S—, whereR is as defined for aryl.

The term “amino” refers to the group —NH₂.

The term “substituted amino” refers to the group —NRR where each R isindependently selected from the group consisting of hydrogen, alkyl,cycloalkyl, carboxyalkyl (for example, benzyloxycarbonyl), aryl,heteroaryl and heterocyclyl provided that both R groups are nothydrogen, or a group —Y—Z, in which Y is optionally substituted alkyleneand Z is alkenyl, cycloalkenyl, or alkynyl. Unless otherwise constrainedby the definition, all substituents may optionally be furthersubstituted by 1-3 substituents chosen from alkyl, carboxy,carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF₃, amino,substituted amino, cyano, and —S(O)_(n)R, where R is alkyl, aryl, orheteroaryl and n is 0, 1 or 2.

The term “carboxyalkyl” refers to the groups —C(O)O-alkyl,

—C(O)O-cycloalkyl, where alkyl and cycloalkyl, are as defined herein,and may be optionally further substituted by alkyl, alkenyl, alkynyl,alkoxy, halogen, CF₃, amino, substituted amino, cyano, or —S(O)_(n)R, inwhich R is alkyl, aryl, or heteroaryl and n is 0, 1 or 2.

The term “cycloalkyl” refers to carbocyclic groups of from 3 to 20carbon atoms having a single cyclic ring or multiple condensed rings.Such cycloalkyl groups include, by way of example, single ringstructures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, andthe like, or multiple ring structures such as adamantanyl,bicyclo[2.2.1]heptane, 1,3,3-trimethylbicyclo[2.2.1]hept-2-yl,(2,3,3-trimethylbicyclo[2.2.1]hept-2-yl), or carbocyclic groups to whichis fused an aryl group, for example indane, and the like.

The term “substituted cycloalkyl” refers to cycloalkyl groups having 1,2, 3, 4 or 5 substituents, and preferably 1, 2, or 3 substituents,selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy,cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy, amino,aminocarbonyl, alkoxycarbonylamino, azido, cyano, halogen, hydroxy,keto, thiocarbonyl, carboxy, carboxyalkyl, arylthio, heteroarylthio,heterocyclylthio, thiol, alkylthio, aryl, aryloxy, heteroaryl,aminosulfonyl, aminocarbonylamino, heteroaryloxy, heterocyclyl,heterocyclooxy, hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-aryl,—SO-heteroaryl, —SO₂-alkyl, SO₂-aryl and —SO₂-heteroaryl. Unlessotherwise constrained by the definition, all substituents may optionallybe further substituted by 1, 2, or 3 substituents chosen from alkyl,carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF₃,amino, substituted amino, cyano, and —S(O)_(n)R, where R is alkyl, aryl,or heteroaryl and n is 0, 1 or 2.

The term “halogen” or “halo” refers to fluoro, bromo, chloro, and iodo.

The term “acyl” denotes a group —C(O)R, in which R is hydrogen,optionally substituted alkyl, optionally substituted cycloalkyl,optionally substituted heterocyclyl, optionally substituted aryl, andoptionally substituted heteroaryl.

The term “heteroaryl” refers to an aromatic cyclic group (i.e., fullyunsaturated) having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15carbon atoms and 1, 2, 3 or 4 heteroatoms selected from oxygen, nitrogenand sulfur within at least one ring. Such heteroaryl groups can have asingle ring (e.g., pyridyl or furyl) or multiple condensed rings (e.g.,indolizinyl, benzothiazolyl, or benzothienyl). Examples of heteroarylsinclude, but are not limited to, [1,2,4]oxadiazole, [1,3,4]oxadiazole,[1,2,4]thiadiazole, [1,3,4]thiadiazole, pyrrole, imidazole, pyrazole,pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole,indole, indazole, purine, quinolizine, isoquinoline, quinoline,phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline,pteridine, carbazole, carboline, phenanthridine, acridine,phenanthroline, isothiazole, phenazine, isoxazole, phenoxazine,phenothiazine, imidazolidine, imidazoline, and the like as well asN-alkoxy-nitrogen containing heteroaryl compounds.

The term “heteroarylene” refers to a diradical of a heteroaryl group asdefined above. This term is exemplified by groups such as2,5-imidazolene, 3,5-[1,2,4]oxadiazolene, 2,4-oxazolene, 1,4-pyrazolene,and the like. For example, 1,4-pyrazolene is:

where A represents the point of attachment.

Unless otherwise constrained by the definition for the heteroaryl orheteroarylene substituent, such heteroaryl or heterarylene groups can beoptionally substituted with 1 to 5 substituents, preferably 1 to 3substituents selected from the group consisting of alkyl, alkenyl,alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy,amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano, halogen,hydroxy, keto, thiocarbonyl, carboxy, carboxyalkyl, arylthio,heteroarylthio, heterocyclylthio, thiol, alkylthio, aryl, aryloxy,heteroaryl, aminosulfonyl, aminocarbonylamino, heteroaryloxy,heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro,—SO-alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl, SO₂-aryl and—SO₂-heteroaryl. Unless otherwise constrained by the definition, allsubstituents may optionally be further substituted by 1-3 substituentschosen from alkyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy,alkoxy, halogen, CF₃, amino, substituted amino, cyano, and —S(O)_(n)R,where R is alkyl, aryl, or heteroaryl and n is 0, 1 or 2.

The term “heteroaralkyl” refers to a heteroaryl group covalently linkedto an alkylene group, where heteroaryl and alkylene are defined herein.“Optionally substituted heteroaralkyl” refers to an optionallysubstituted heteroaryl group covalently linked to an optionallysubstituted alkylene group. Such heteroaralkyl groups are exemplified by3-pyridylmethyl, quinolin-8-ylethyl, 4-methoxythiazol-2-ylpropyl, andthe like.

The term “heteroaryloxy” refers to the group heteroaryl-O—.

The term “heterocyclyl” refers to a monoradical saturated or partiallyunsaturated group having a single ring or multiple condensed rings,having from 1 to 40 carbon atoms and from 1 to 10 hetero atoms,preferably 1, 2, 3 or 4 heteroatoms, selected from nitrogen, sulfur,phosphorus, and/or oxygen within the ring. Heterocyclic groups can havea single ring or multiple condensed rings, and includetetrahydrofuranyl, morpholino, piperidinyl, piperazino, dihydropyridino,and the like.

Unless otherwise constrained by the definition for the heterocyclicsubstituent, such heterocyclic groups can be optionally substituted with1, 2, 3, 4 or 5, and preferably 1, 2 or 3 substituents, selected fromthe group consisting of alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl,cycloalkenyl, acyl, acylamino, acyloxy, amino, aminocarbonyl,alkoxycarbonylamino, azido, cyano, halogen, hydroxy, keto, thiocarbonyl,carboxy, carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio,thiol, alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl,aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-aryl, —SO-heteroaryl,—SO₂-alkyl, SO₂-aryl and —SO₂-heteroaryl. Unless otherwise constrainedby the definition, all substituents may optionally be furthersubstituted by 1-3 substituents chosen from alkyl, carboxy,carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF₃, amino,substituted amino, cyano, and —S(O)_(n)R, where R is alkyl, aryl, orheteroaryl and n is 0, 1 or 2.

The term “thiol” refers to the group —SH.

The term “substituted alkylthio” refers to the group —S-substitutedalkyl.

The term “heteroarylthiol” refers to the group —S-heteroaryl wherein theheteroaryl group is as defined above including optionally substitutedheteroaryl groups as also defined above.

The term “sulfoxide” refers to a group —S(O)R, in which R is alkyl,aryl, or heteroaryl. “Substituted sulfoxide” refers to a group —S(O)R,in which R is substituted alkyl, substituted aryl, or substitutedheteroaryl, as defined herein.

The term “sulfone” refers to a group —S(O)₂R, in which R is alkyl, aryl,or heteroaryl. “Substituted sulfone” refers to a group —S(O)₂R, in whichR is substituted alkyl, substituted aryl, or substituted heteroaryl, asdefined herein.

The term “keto” refers to a group —C(O)—. The term “thiocarbonyl” refersto a group —C(S)—. The term “carboxy” refers to a group —C(O)—OH.

“Optional” or “optionally” means that the subsequently described eventor circumstance may or may not occur, and that the description includesinstances where said event or circumstance occurs and instances in whichit does not.

The term “compound of Formula I” is intended to encompass the compoundsof the invention as disclosed, and the pharmaceutically acceptablesalts, pharmaceutically acceptable esters, prodrugs, hydrates andpolymorphs of such compounds. Additionally, the compounds of theinvention may possess one or more asymmetric centers, and can beproduced as a racemic mixture or as individual enantiomers ordiastereoisomers. The number of stereoisomers present in any givencompound of Formula I depends upon the number of asymmetric centerspresent (there are 2^(n) stereoisomers possible where n is the number ofasymmetric centers). The individual stereoisomers may be obtained byresolving a racemic or non-racemic mixture of an intermediate at someappropriate stage of the synthesis, or by resolution of the compound ofFormula I by conventional means. The individual stereoisomers (includingindividual enantiomers and diastereoisomers) as well as racemic andnon-racemic mixtures of stereoisomers are encompassed within the scopeof the present invention, all of which are intended to be depicted bythe structures of this specification unless otherwise specificallyindicated.

“Isomers” are different compounds that have the same molecular formula.

“Stereoisomers” are isomers that differ only in the way the atoms arearranged in space.

“Enantiomers” are a pair of stereoisomers that are non-superimposablemirror images of each other. A 1:1 mixture of a pair of enantiomers is a“racemic” mixture. The term “(±)” is used to designate a racemic mixturewhere appropriate.

“Diastereoisomers” are stereoisomers that have at least two asymmetricatoms, but which are not mirror-images of each other.

The absolute stereochemistry is specified according to theCahn-Ingold-Prelog R-S system. When the compound is a pure enantiomerthe stereochemistry at each chiral carbon may be specified by either Ror S. Resolved compounds whose absolute configuration is unknown aredesignated (+) or (−) depending on the direction (dextro- orlaevorotary) which they rotate the plane of polarized light at thewavelength of the sodium D line.

The term “therapeutically effective amount” refers to that amount of acompound of Formula I that is sufficient to effect treatment, as definedbelow, when administered to a mammal in need of such treatment. Thetherapeutically effective amount will vary depending upon the subjectand disease condition being treated, the weight and age of the subject,the severity of the disease condition, the manner of administration andthe like, which can readily be determined by one of ordinary skill inthe art.

The term “treatment” or “treating” means any treatment of a disease in amammal, including:

-   -   (i) preventing the disease, that is, causing the clinical        symptoms of the disease not to develop;    -   (ii) inhibiting the disease, that is, arresting the development        of clinical symptoms; and/or    -   (iii) relieving the disease, that is, causing the regression of        clinical symptoms.

In many cases, the compounds of this invention are capable of formingacid and/or base salts by virtue of the presence of amino and/orcarboxyl groups or groups similar thereto. The term “pharmaceuticallyacceptable salt” refers to salts that retain the biologicaleffectiveness and properties of the compounds of Formula I, and whichare not biologically or otherwise undesirable. Pharmaceuticallyacceptable base addition salts can be prepared from inorganic andorganic bases. Salts derived from inorganic bases, include by way ofexample only, sodium, potassium, lithium, ammonium, calcium andmagnesium salts. Salts derived from organic bases include, but are notlimited to, salts of primary, secondary and tertiary amines, such asalkyl amines, dialkyl amines, trialkyl amines, substituted alkyl amines,di(substituted alkyl)amines, tri(substituted alkyl)amines, alkenylamines, dialkenyl amines, trialkenyl amines, substituted alkenyl amines,di(substituted alkenyl)amines, tri(substituted alkenyl)amines,cycloalkyl amines, di(cycloalkyl)amines, tri(cycloalkyl)amines,substituted cycloalkyl amines, disubstituted cycloalkyl amine,trisubstituted cycloalkyl amines, cycloalkenyl amines,di(cycloalkenyl)amines, tri(cycloalkenyl)amines, substitutedcycloalkenyl amines, disubstituted cycloalkenyl amine, trisubstitutedcycloalkenyl amines, aryl amines, diaryl amines, triaryl amines,heteroaryl amines, diheteroaryl amines, triheteroaryl amines,heterocyclic amines, diheterocyclic amines, triheterocyclic amines,mixed di- and tri-amines where at least two of the substituents on theamine are different and are selected from the group consisting of alkyl,substituted alkyl, alkenyl, substituted alkenyl, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl,heterocyclic, and the like. Also included are amines where the two orthree substituents, together with the amino nitrogen, form aheterocyclic or heteroaryl group.

Specific examples of suitable amines include, by way of example only,isopropylamine, trimethyl amine, diethyl amine, tri(iso-propyl)amine,tri(n-propyl)amine, ethanolamine, 2-dimethylaminoethanol, tromethamine,lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline,betaine, ethylenediamine, glucosamine, N-alkylglucamines, theobromine,purines, piperazine, piperidine, morpholine, N-ethylpiperidine, and thelike.

Pharmaceutically acceptable acid addition salts may be prepared frominorganic and organic acids. Salts derived from inorganic acids includehydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, and the like. Salts derived from organic acids includeacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid,malic acid, malonic acid, succinic acid, maleic acid, fumaric acid,tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, p-toluene-sulfonic acid,salicylic acid, and the like.

As used herein, “pharmaceutically acceptable carrier” includes any andall solvents, dispersion media, coatings, antibacterial and antifungalagents, isotonic and absorption delaying agents and the like. The use ofsuch media and agents for pharmaceutically active substances is wellknown in the art. Except insofar as any conventional media or agent isincompatible with the active ingredient, its use in the therapeuticcompositions is contemplated. Supplementary active ingredients can alsobe incorporated into the compositions.

As used herein, the term “prodrug” denotes a compound that ismetabolized in-vivo to a compound that is active as an A_(2B) adenosinereceptor antagonist.

Nomenclature

The naming and numbering of the compounds of the invention isillustrated with a representative compound of Formula I in which R¹ isn-propyl, R² is ethyl, R⁴ is 3-trifluorophenyl, X is hydrogen, and Y is—C(O)CH₂CH₂CH₃;

which is named:

-   [3-ethyl-2,6-dioxo-l-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl    butanoate.

Synthetic Reaction Parameters

The terms “solvent”, “inert organic solvent” or “inert solvent” mean asolvent inert under the conditions of the reaction being described inconjunction therewith [including, for example, benzene, toluene,acetonitrile, tetrahydrofuran (“THF”), dimethylformamide (“DMF”),chloroform, methylene chloride (or dichloromethane), diethyl ether,methanol, pyridine and the like]. Unless specified to the contrary, thesolvents used in the reactions of the present invention are inertorganic solvents. The term “q.s.” means adding a quantity sufficient toachieve a stated function, e.g., to bring a solution to the desiredvolume (i.e., 100%).

Synthesis of the Compounds of Formula I

A method for preparing compounds of Formula I in which Y is optionallysubstituted lower alkyl, optionally substituted aryl, or optionallysubstituted heteroaryl is shown in Reaction Scheme I.

where R¹, R²,R⁴, X and Y are as defined above.

In general, the compound of formula (1) is reacted in a polar solvent,for example N,N-dimethylformamide, with a compound of formula YOCHXCl.The reaction is carried out at a temperature of about 30 to 80° C.,preferably about 60° C., in the presence of a base, preferably aninorganic base, for example potassium carbonate, for about 8-24 hours.When the reaction is substantially complete, the product of Formula I isisolated by conventional means, for example preparative chromatography.

The starting compound of formula (1) can be prepared by those techniquesdisclosed in U.S. Pat. No. 6,825,349, or those disclosed in U.S. patentapplication Ser. No.10/719,102, publication number 20040176399, theentire contents of which are hereby incorporated by reference.

When Y is —C(O)R, in which R is a heterocycle, the compound of formula(2) (RC(O)OCHXCl) is either commercially available or can be prepared asshown below, using pyridine as an example.

In general, the carboxylic acid of formula (a) is reacted in an inertsolvent, for example dichloromethane, with a chloromethyl derivative offormula (b) in the presence of a quarternary salt, for exampletetrabutylammonium sulfate. The reaction is carried out at a temperatureof about 0° C., in the presence of a base, preferably an inorganic base,for example sodium bicarbonate, followed by reaction at room temperaturefor about 2-10 hours. When the reaction is substantially complete, theproduct, chloromethyl pyridine-3-carboxylate, is isolated byconventional means.

Carbamate derivatives can be prepared as shown in Reaction Scheme II.

where R¹, R² and R⁴, are as defined above, and R_(a)R_(b)NH representsan amine.

In general, the amine of formula R_(a)R_(b)NH is reacted in a polarsolvent, for example N,N-dimethylformamide, with chloromethylchloroformate at a temperature of about 0° C., in the presence of abase, preferably an inorganic base, for example potassium carbonate, forabout 1 hour. Then a solution of the compound of formula (1) in a polarsolvent at 0° C. is added, and the mixture reacted for 24 hours,allowing the temperature to rise to room temperature. When the reactionis substantially complete, the product of Formula I is isolated byconventional means, for example preparative chromatography.

To prepare an ether derivative of a compound of formula (1), thecompound of formula (1) is reacted conventionally with an appropriatechloromethyl ether.

A method for preparing compounds of Formula I in which Y is —P(O)(OH)₂is shown in Reaction Scheme III.

Reaction Scheme III

Step 1

In general, the compound of formula (2) is reacted with a compound offormula (1) in a polar solvent, for example N,N-dimethylformamide, at atemperature of about 30-90° C., in the presence of a base, preferably aninorganic base, for example potassium carbonate, for about 4-24 hours.When the reaction is substantially complete, the product of formula (3)is isolated by conventional means and purified, for example preparativechromatography.

Step 2

The product of formula (3) is deprotected conventionally with a strongacid, for example trifluoroacetic acid, or alternatively a weak acidsuch as formic acid, in an inert solvent, for example dichloromethane.The reaction is conducted at about room temperature for about 4-24hours. When the reaction is substantially complete, the product ofFormula I in which Y is —P(O)(OH)₂ is isolated by conventional means andpurified, for example preparative chromatography.

Starting Material of Formula (2)

The compound of formula (2), di-tert-butyl chloromethyl phosphate, isprepared from bis(tert-butoxy)phosphino-1-ol as shown below.

Step 1

In general, the compound of formula (a), bis(tert-butoxy)phosphino-1-ol,is reacted with an oxidizing, for example potassium permanganate, in thepresence of a mild base, for example potassium bicarbonate, in anaqueous solvent. The reaction is initially conducted at a temperature ofabout 0° C., and then at about room temperature for about 1 hour. Whenthe reaction is substantially complete, the product of formula (b),ditert-butyl hydrogen phosphate, is isolated by conventional means, forexample by acidification and filtration of the phosphate thus formed.

Step 2

Initially a tetramethylammonium salt of (b) is prepared by reaction ofditert-butyl hydrogen phosphate with tetramethylammonium hydroxide in aninert solvent, for example acetone, at a temperature of about 0° C. Thetetramethylammonium salt of ditert-butyl hydrogen phosphate is isolatedby conventional means, for example by removal of the solvent.

The tetramethylammonium salt of ditert-butyl hydrogen phosphate is thenreacted with a dihalomethane derivative, for example dibromomethane orchloroiodomethane, in an inert solvent, for example 1,2-dimethoxyethane.The reaction is conducted at a temperature of about 60-90° C. When thereaction is substantially complete, the product of formula (2) isisolated by conventional means.

Utility, Testing and Administration General Utility

The compounds of Formula I are effective in-vivo for the treatment ofconditions that respond to administration of A_(2B) adenosine receptorantagonists. Such conditions include, but are not limited to, at leastone of diarrhea, atherosclerosis, restenosis, diabetic retinopathy,cancer, senile dementia, Alzheimer's disease, Parkinson's disease,traumatic brain injury, and Type I hypersensitivity reactions, includingchronic obstructive pulmonary disease (COPD), asthma, atopic eczema, andhay fever.

Pharmaceutical Compositions

The compounds of Formula I are usually administered in the form ofpharmaceutical compositions. This invention therefore providespharmaceutical compositions that contain, as the active ingredient, oneor more of the compounds of Formula I, or a pharmaceutically acceptablesalt or ester thereof, and one or more pharmaceutically acceptableexcipients, carriers, including inert solid diluents and fillers,diluents, including sterile aqueous solution and various organicsolvents, permeation enhancers, solubilizers and adjuvants. Thecompounds of Formula I may be administered alone or in combination withother therapeutic agents. Such compositions are prepared in a mannerwell known in the pharmaceutical art (see, e.g., Remington'sPharmaceutical Sciences, Mace Publishing Co., Philadelphia, Pa. 17^(th)Ed. (1985) and “Modern Pharmaceutics”, Marcel Dekker, Inc. 3^(rd) Ed.(G. S. Banker & C. T. Rhodes, Eds.).

Administration

The compounds of Formula I may be administered in either single ormultiple doses by any of the accepted modes of administration of agentshaving similar utilities, for example as described in those patents andpatent applications incorporated by reference, including rectal, buccal,intranasal and transdermal routes, by intra-arterial injection,intravenously, intraperitoneally, parenterally, intramuscularly,subcutaneously, orally, topically, as an inhalant, or via an impregnatedor coated device such as a stent, for example, or an artery-insertedcylindrical polymer.

One mode for administration is parental, particularly by injection. Theforms in which the novel compositions of the present invention may beincorporated for administration by injection include aqueous or oilsuspensions, or emulsions, with sesame oil, corn oil, cottonseed oil, orpeanut oil, as well as elixirs, mannitol, dextrose, or a sterile aqueoussolution, and similar pharmaceutical vehicles. Aqueous solutions insaline are also conventionally used for injection, but less preferred inthe context of the present invention. Ethanol, glycerol, propyleneglycol, liquid polyethylene glycol, and the like (and suitable mixturesthereof), cyclodextrin derivatives, and vegetable oils may also beemployed. The proper fluidity can be maintained, for example, by the useof a coating, such as lecithin, by the maintenance of the requiredparticle size in the case of dispersion and by the use of surfactants.The prevention of the action of microorganisms can be brought about byvarious antibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, sorbic acid, thimerosal, and the like.

Sterile injectable solutions are prepared by incorporating the compoundof Formula I in the required amount in the appropriate solvent withvarious other ingredients as enumerated above, as required, followed byfiltered sterilization. Generally, dispersions are prepared byincorporating the various sterilized active ingredients into a sterilevehicle which contains the basic dispersion medium and the requiredother ingredients from those enumerated above. In the case of sterilepowders for the preparation of sterile injectable solutions, thepreferred methods of preparation are vacuum-drying and freeze-dryingtechniques which yield a powder of the active ingredient plus anyadditional desired ingredient from a previously sterile-filteredsolution thereof.

Oral administration is another route for administration of the compoundsof Formula I. Administration may be via capsule or enteric coatedtablets, or the like. In making the pharmaceutical compositions thatinclude at least one compound of Formula I, the active ingredient isusually diluted by an excipient and/or enclosed within such a carrierthat can be in the form of a capsule, sachet, paper or other container.When the excipient serves as a diluent, in can be a solid, semi-solid,or liquid material (as above), which acts as a vehicle, carrier ormedium for the active ingredient. Thus, the compositions can be in theform of tablets, pills, powders, lozenges, sachets, cachets, elixirs,suspensions, emulsions, solutions, syrups, aerosols (as a solid or in aliquid medium), ointments containing, for example, up to 10% by weightof the active compound, soft and hard gelatin capsules, sterileinjectable solutions, and sterile packaged powders.

Some examples of suitable excipients include lactose, dextrose, sucrose,sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates,tragacanth, gelatin, calcium silicate, microcrystalline cellulose,polyvinylpyrrolidone, cellulose, sterile water, syrup, and methylcellulose. The formulations can additionally include: lubricating agentssuch as talc, magnesium stearate, and mineral oil; wetting agents;emulsifying and suspending agents; preserving agents such as methyl- andpropylhydroxy-benzoates; sweetening agents; and flavoring agents.

The compositions of the invention can be formulated so as to providequick, sustained or delayed release of the active ingredient afteradministration to the patient by employing procedures known in the art.Controlled release drug delivery systems for oral administration includeosmotic pump systems and dissolutional systems containing polymer-coatedreservoirs or drug-polymer matrix formulations. Examples of controlledrelease systems are given in U.S. Pat. Nos. 3,845,770; 4,326,525;4,902514; and 5,616,345. Another formulation for use in the methods ofthe present invention employs transdermal delivery devices (“patches”).Such transdermal patches may be used to provide continuous ordiscontinuous infusion of the compounds of the present invention incontrolled amounts. The construction and use of transdermal patches forthe delivery of pharmaceutical agents is well known in the art. See,e.g., U.S. Pat. Nos. 5,023,252, 4,992,445 and 5,001,139. Such patchesmay be constructed for continuous, pulsatile, or on demand delivery ofpharmaceutical agents.

The compositions are preferably formulated in a unit dosage form. Theterm “unit dosage forms” refers to physically discrete units suitable asunitary dosages for human subjects and other mammals, each unitcontaining a predetermined quantity of active material calculated toproduce the desired therapeutic effect, in association with a suitablepharmaceutical excipient (e.g., a tablet, capsule, ampoule). Thecompounds of Formula I are effective over a wide dosage range and isgenerally administered in a pharmaceutically effective amount.Preferably, for oral administration, each dosage unit contains from 10mg to 2 g of a compound of Formula I, more preferably from 10 to 700 mg,and for parenteral administration, preferably from 10 to 700 mg of acompound of Formula I, more preferably about 50-200 mg. It will beunderstood, however, that the amount of the compound of Formula Iactually administered will be determined by a physician, in the light ofthe relevant circumstances, including the condition to be treated, thechosen route of administration, the actual compound administered and itsrelative activity, the age, weight, and response of the individualpatient, the severity of the patient's symptoms, and the like.

For preparing solid compositions such as tablets, the principal activeingredient is mixed with a pharmaceutical excipient to form a solidpreformulation composition containing a homogeneous mixture of acompound of the present invention. When referring to thesepreformulation compositions as homogeneous, it is meant that the activeingredient is dispersed evenly throughout the composition so that thecomposition may be readily subdivided into equally effective unit dosageforms such as tablets, pills and capsules.

The tablets or pills of the present invention may be coated or otherwisecompounded to provide a dosage form affording the advantage of prolongedaction, or to protect from the acid conditions of the stomach. Forexample, the tablet or pill can comprise an inner dosage and an outerdosage component, the latter being in the form of an envelope over theformer. The two components can be separated by an enteric layer thatserves to resist disintegration in the stomach and permit the innercomponent to pass intact into the duodenum or to be delayed in release.A variety of materials can be used for such enteric layers or coatings,such materials including a number of polymeric acids and mixtures ofpolymeric acids with such materials as shellac, cetyl alcohol, andcellulose acetate.

Compositions for inhalation or insufflation include solutions andsuspensions in pharmaceutically acceptable, aqueous or organic solvents,or mixtures thereof, and powders. The liquid or solid compositions maycontain suitable pharmaceutically acceptable excipients as describedsupra. Preferably the compositions are administered by the oral or nasalrespiratory route for local or systemic effect. Compositions inpreferably pharmaceutically acceptable solvents may be nebulized by useof inert gases. Nebulized solutions may be inhaled directly from thenebulizing device or the nebulizing device may be attached to a facemask tent, or intermittent positive pressure breathing machine.Solution, suspension, or powder compositions may be administered,preferably orally or nasally, from devices that deliver the formulationin an appropriate manner.

The following examples are included to demonstrate preferred embodimentsof the invention. It should be appreciated by those of skill in the artthat the techniques disclosed in the examples which follow representtechniques discovered by the inventor to function well in the practiceof the invention, and thus can be considered to constitute preferredmodes for its practice. However, those of skill in the art should, inlight of the present disclosure, appreciate that many changes can bemade in the specific embodiments which are disclosed and still obtain alike or similar result without departing from the spirit and scope ofthe invention.

EXAMPLE 1 Preparation of a Compound of Formula I Preparation of aCompound of Formula I where R¹ is n-Propyl, R² is Ethyl, R⁴ is3-Trifluoromethylphenyl, X is Hydrogen, and Y is n-Butanoyl

To a solution of3-ethyl-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}-pyrazol-4-yl)-1,3,7-trihydropurine-2,6-dione(250 mg, 0.56 mmol) in N,N-dimethylformamide (10 ml) was added potassiumcarbonate (230 mg, 1.68 mmol), followed by chloromethylbutyrate (230 mg,1.68 mmol), and the mixture was stirred at 60° C. for 16 hours. Thesolid was filtered off, and solvent removed from the filtrate underreduced pressure. The residue was chromatographed on silica gel, elutingwith 30% ethyl acetate/hexane, yielding[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methylbutanoate (150 mg). ¹H NMR (CDCl₃): d 7.98 (s, 1H), 7.97 (s, 1H),7.65-7.45 (m, 4H), 6.35 (s, 2H), 5.44 (s, 2H), 4.19 (q, J =8 Hz, 2H),3.98 (q, J=2 Hz, 2H), 2.33 (t, J=8 Hz, 2H), 1.75-1.60 (m, 4H), 1.36 (t,J=8 Hz, 3H), 0.96 (t, J =8 Hz, 3 H), 0.92 (t, J=8 Hz, 3H).

B. Preparation of other Compounds of Formula I

Similarly, following the procedure of 1A above, but replacingchloromethylbutyrate by chloromethyl-2,2-dimethylpropanoate,[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl-2,2-dimethylpropanoatewas prepared. ¹H NMR (CDCl₃): d 7.98 (s, 1H), 7.97 (s, 1H), 7.65-7.45(m, 4H), 6.33 (s, 2H), 5.43 (s, 2H), 4.19 (q, J=8 Hz, 2H), 3.98 (q, J=2Hz, 2H), 1.75-1.64 (m, 2H), 1.37 (t, J =8 Hz, 3H), 1.16 (s, 9H), 0.96(t, J=8 Hz, 3H).

Similarly, following the procedure of 1A above, but replacingchloromethylbutyrate by chloromethyl acetate,[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methylacetate was prepared. ¹H NMR (CDCl₃): d 7.98 (s, 1H), 7.97 (s, 1H),7.65-7.45 (m, 4H), 6.35 (s, 2H), 5.44 (s, 2H), 4.19 (q, J=8 Hz, 2H),3.98 (q, J =2 Hz, 2H), 2.33 (t, J=8 Hz, 2H), 1.75-1.60 (m, 4H), 1.36 (t,J =8 Hz, 3H), 0.96 (t, J=8 Hz, 3H), 0.92 (t, J=8 Hz, 3H),

Similarly, following the procedure of 1A above, but replacingchloromethylbutyrate bychloro(2S)-1-[benzyloxycarbonyl]pyrrolidine-2-carboxylate,[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl(2S)-1-[benzyloxycarbonyl]pyrrolidine-2-carboxylate was prepared. TheNMR of this compound was satisfactory.

C. Preparation of other Compounds of Formula I

Similarly, following the procedure of 1A above, but replacingchloromethylbutyrate by other compounds of the formula YOCHXCl, in whichX and Y are as defined above, the following compounds of Formula I areprepared:

-   [3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl    2-methylpropanoate;-   [3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethy)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl    benzoate;-   [3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethy)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl    3-(trifluoromethyl)benzoate;-   [3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethy)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl    2-phenylacetate;-   [3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethy)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]ethyl    butanoate;-   [3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl    propanoate;-   [3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl    pentanoate;-   [3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethy)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl    hexanoate;-   [3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethy)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl    octanoate;-   [3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethy)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl    3-methylbutanoate;-   [3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethy)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl    cyclopentanecarboxylate;-   [3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethy)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl    cyclohexanecarboxylate;-   2-({[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethy)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl}oxycarbonyl)acetic    acid;-   3-({[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethy)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl}oxycarbonyl)propanoic    acid;-   [3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethy)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl    3-methoxypropanoate;-   [3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethy)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl    3-hydroxybutanoate;-   [3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethy)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl    3-(oxyphosphinyloxyphosphinyl)butanoate;-   [3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl    3-[(oxyphosphinyloxyphosphinyemethoxy]butanoate;-   [3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl    benzoate;-   [3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl    4-piperazinylbenzoate;-   [3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethy)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl    4-morpholin-4-ylbenzoate; and-   [3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethy)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl    pyridine-3-carboxylate.

EXAMPLE 2 Preparation of a Carbamate Derivative of a Compound of Formula(1) Preparation of[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[13-(trifluoromethyl)phenyl]methyl}-pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl4-methylpiperazinecarboxylate

Chloromethyl chloroformate (0.319 mmol) and 1-methylpiperazine (0.319mmol) were mixed in N,N-dimethylformamide (2 ml) at 0° C. in thepresence of potassium carbonate (1.325 mmol). After 1 hour, a precooledsolution of3-ethyl-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}-pyrazol-4-yl)-1,3,7-trihydropurine-2,6-dione(0.265 mmol) in N,N-dimethylformamide (1 ml) was added, and the mixturewas stirred for 24 hours, allowing the temperature to rise to roomtemperature. Solvent was removed under reduced pressure, and the residueapplied to a preparative thin layer chromatography plate, eluting with5% methanol/methylene chloride, providing 150 mg of[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl4-methylpiperazinecarboxylate.

Similarly, the following compounds were prepared:

-   N-[2-(dimethylamino)ethyl]{[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)(1,3,7-trihydropurin-7-yl)]methoxy}-carboxamide;-   N-[2-(dimethylamino)ethyl]{[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)(1,3,7-trihydropurin-7-yl)]methoxy}-N-methylcarboxamide;    and-   N-[((2S)-1-ethyl(2-piperidyl))methyl]{[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)(1,3,7-trihydropurin-7-yl)]methoxy}carboxamide.

EXAMPLE 3 Preparation of a Phosphate Derivative of a Compound of Formula(1) Preparation of[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}-pyrazol-4-yl)(1,3,7-trihydropurin-7-yl)]methyldihydrogen phosphate

Step 1—Preparation of Di-Tert-Butyl Chloromethyl Phosphate (Formula (2))

a) Preparation of Ditert-Butyl Hydrogen Phosphate

To a stirred solution of bis(tert-butoxy)phosphino-1-ol (0.78 g, 4 mmol)and potassium bicarbonate (0.6 g, 2.4 mmol) in water (4 ml) at 0° C. wasadded (in portions) potassium permanganate (0.44 g, 2.8 mmol). Themixture was allowed to warm to room temperature, and stirred for 1 hour.Decolorizing charcoal (60 mg) was then added, and the mixture stirred at60° C. for 15 minutes, and then filtered. The solid thus obtained waswashed with water (30 ml), and the combined filtrates were treated witha further 100 mg of decolorizing charcoal at 60° C. for 20 minutes. Themixture was filtered, and the filtrate cooled to 0° C. and carefullyacidified with concentrated hydrochloric acid (2 ml) with stirring. Theprecipitate was filtered off, washed with cold water, to provideditert-butyl hydrogen phosphate as a white solid.

Preparation of the Tetramethylammonium Salt of Ditert-Butyl HydrogenPhosphate

A solution of the di-tert-butyl hydrogen phosphate obtained in step a)was dissolved in acetone (10 ml) and cooled to 0° C. To this solutionwas added a 10% aqueous solution of tetramethylammonium hydroxide (2.4ml, 2.6 mmol), and the homogeneous solution was evaporated under reducedpressure to provide a solid, which was crystallized from refluxing1,2-dimethoxyethane to provide tetramethylammonium ditert-butyl hydrogenphosphate as a white solid.

The tetramethylammonium ditert-butyl hydrogen phosphate obtained in stepb was dissolved in refluxing 1,2-dimethoxymethane (15 ml), andchloroiodomethane (3.2 g, 18.1 mmol) added, and the mixture was refluxedfor 90 minutes. The solvent was removed under reduced pressure, and theresidue, di-tert-butyl chloromethyl phosphate, was used as such withoutfurther purification.

Step 2

A solution of3-ethyl-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}-pyrazol-4-yl)-1,3,7-trihydropurine-2,6-dione(0.47 g, 1 mmol) was dissolved in 20 ml of N,N-dimethylformamide, andpotassium carbonate (0.42 g, 4 mmol) was added, followed bydi-tert-butyl chloromethyl phosphate (0.34 g, 1.32 mmol), and themixture was stirred at 60° C. overnight. The reaction mixture wascooled, and the precipitate filtered off, washing with ethyl acetate.The filtrate was concentrated under reduced pressure, and the residuewas purified by preparative thin layer chromatography, eluting with 4%methanol/methylene chloride, to provide tert-butyl[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)(1,3,7-trihydropurin-7-yl)]methylmethylethyl phosphate(0.26 g) as a colorless oil.

Step 3

A solution of tert-butyl[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)(1,3,7-trihydropurin-7-yl)]methylmethylethyl phosphate (80 mg, 0.12 mmol) was dissolved in methylenechloride (6 ml) and trifluoroacetic acid (0.72 mmol) was added. Themixture was stirred at room temperature overnight. The solvent wasremoved under reduced pressure, and the solid white residue wastriturated with ether and collected by filtration, providing[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}-pyrazol-4-yl)(1,3,7-trihydropurin-7-yl)]methyldihydrogen phosphate (41 mg).

NMR ¹H-NMR (DMSO-d6) δ 8.70 (s, 1H), 8.15 (s, 1H), 7.74 (s, 1H),7.69-7.71 (m, 1H), 7.60-7.63 (m, 2H), 6.12 (d, 2H, J =5.4 Hz), 5.54 (s,2H), 4.06 (q, 2H, J=13.8 Hz), 3.84 (t, 2H, J=7.4 Hz), 1.52-1.62 (m, 2H),1.25 (t, 3H, J=7.0 Hz), 0.87 (t, 3H), J=7.4 Hz); MS m/z 579.02 (M⁺+Na)

EXAMPLE 4 Preparation of a Compound of Formula (1) Preparation of aCompound of Formula Tin which R¹ is n-Propyl, R² is Ethyl, R⁴ is3-Trifluoromethylphenyl, X is Hydrogen, and Y is n-Butanoyl A.Preparation of chloromethyl pyridine-3-carboxylate

A mixture of nicotinic acid (200 mg, 1.6 mmol), sodium bicarbonate (540mg, 6.4 mmol), and tetrabutylammonium sulfate (54 mg, 0.16 mmol) wasdissolved in a mixture of 4 ml of dichloromethane and 4 ml of water, andcooled to 0° C. To this stirred mixture was addedchloromethylchlorosulfone (165 μl, 1.6 mmol) in 1 ml of dichloromethane,and the mixture was allowed to warm to room temperature, stirringovernight. The organic layer was separated, washed with brine, driedover sodium sulfate, and concentrated under reduced pressure to a yellowoil, which was dissolved in dichloromethane and filtered through asilica plug. Removal of the solvent under reduced pressure providedchloromethyl pyridine-3-carboxylate (70 mg).

B. Preparation of[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]-methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methylpyridine-3-carboxylate

A solution of3-ethyl-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}-pyrazol-4-yl)-1,3,7-trihydropurine-2,6-dione(200 mg, 0.43 mmol) was dissolved in 2 ml of N,N-dimethylformamide, andpotassium carbonate (120 mg, 0.86 mmol) was added, followed bychloromethyl pyridine-3-carboxylate (220 mg, 1.3 mmol). The mixture wasstirred at 60° C. overnight, the solid material filtered off, and thefiltrate evaporated under reduced pressure. The residue was purified bythin layer chromatography, eluting with 5% methanol/dichloromethane, toprovide[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methylpyridine-3-carboxylate (66 mg).

NMR of the product was satisfactory.

EXAMPLE 5

Hard gelatin capsules containing the following ingredients are prepared:

Quantity Ingredient (mg/capsule) Active Ingredient 30.0 Starch 305.0Magnesium stearate 5.0

The above ingredients are mixed and filled into hard gelatin capsules.

EXAMPLE 6

A tablet formula is prepared using the ingredients below:

Quantity Ingredient (mg/tablet) Active Ingredient 25.0 Cellulose,microcrystalline 200.0 Colloidal silicon dioxide 10.0 Stearic acid 5.0The components are blended and compressed to form tablets.

EXAMPLE 7

A dry powder inhaler formulation is prepared containing the followingcomponents:

Ingredient Weight % Active Ingredient 5 Lactose 95The active ingredient is mixed with the lactose and the mixture is addedto a dry powder inhaling appliance.

EXAMPLE 8

Tablets, each containing 30 mg of active ingredient, are prepared asfollows:

Quantity Ingredient (mg/tablet) Active Ingredient 30.0 mg Starch 45.0 mgMicrocrystalline cellulose 35.0 mg Polyvinylpyrrolidone 4.0 mg (as 10%solution in sterile water) Sodium carboxymethyl starch 4.5 mg Magnesiumstearate 0.5 mg Talc 1.0 mg Total 120 mg

The active ingredient, starch and cellulose are passed through a No. 20mesh U.S. sieve and mixed thoroughly. The solution ofpolyvinylpyrrolidone is mixed with the resultant powders, which are thenpassed through a 16 mesh U.S. sieve. The granules so produced are driedat 50° C. to 60° C. and passed through a 16 mesh U.S. sieve. The sodiumcarboxymethyl starch, magnesium stearate, and talc, previously passedthrough a No. 30 mesh U.S. sieve, are then added to the granules which,after mixing, are compressed on a tablet machine to yield tablets eachweighing 120 mg.

EXAMPLE 9

Suppositories, each containing 25 mg of active ingredient are made asfollows:

Ingredient Amount Active Ingredient   25 mg Saturated fatty acidglycerides to 2,000 mg

The active ingredient is passed through a No. 60 mesh U.S. sieve andsuspended in the saturated fatty acid glycerides previously melted usingthe minimum heat necessary. The mixture is then poured into asuppository mold of nominal 2.0 g capacity and allowed to cool.

EXAMPLE 10

Suspensions, each containing 50 mg of active ingredient per 5.0 mL doseare made as follows:

Ingredient Amount Active Ingredient 50.0 mg Xanthan gum 4.0 mg Sodiumcarboxymethyl cellulose (11%) 50.0 mg Microcrystalline cellulose (89%)Sucrose 1.75 g Sodium benzoate 10.0 mg Flavor and Color q.v. Purifiedwater to 5.0 mL

The active ingredient, sucrose and xanthan gum are blended, passedthrough a No. 10 mesh U.S. sieve, and then mixed with a previously madesolution of the microcrystalline cellulose and sodium carboxymethylcellulose in water. The sodium benzoate, flavor, and color are dilutedwith some of the water and added with stirring. Sufficient water is thenadded to produce the required volume.

EXAMPLE 11

A subcutaneous formulation may be prepared as follows:

Ingredient Quantity Active Ingredient 5.0 mg Corn Oil 1.0 mL

EXAMPLE 12

An injectable preparation is prepared having the following composition:

Ingredients Amount Active ingredient 2.0 mg/ml Mannitol, USP  50 mg/mlGluconic acid, USP q.s. (pH 5-6) water (distilled, sterile) q.s. to 1.0ml Nitrogen Gas, NF q.s.

EXAMPLE 13

A topical preparation is prepared having the following composition:

Ingredients grams Active ingredient 0.2-10 Span 60 2.0 Tween 60 2.0Mineral oil 5.0 Petrolatum 0.10 Methyl paraben 0.15 Propyl paraben 0.05BHA (butylated hydroxy anisole) 0.01 Water q.s. to 100

All of the above ingredients, except water, are combined and heated to60° C. with stirring. A sufficient quantity of water at 60° C. is thenadded with vigorous stirring to emulsify the ingredients, and water thenadded q.s. 100 g.

EXAMPLE 14 A_(2B) Adenosine Receptor Assays Methods Radioligand Bindingfor A_(2B) Adenosine Receptor.

Human A_(2B) adenosine receptor cDNA is stably transfected into HEK-293cells (referred to as HEK-A2B cells). Monolayer of HEK-A2B cells arewashed with PBS once and harvested in a buffer containing 10 mM HEPES(pH 7.4), 10 mM EDTA and protease inhibitors. These cells arehomogenized in polytron for 1 minute at setting 4 and centrifuged at29000 g for 15 minutes at 4° C. The cell pellets are washed once with abuffer containing 10 mM HEPES (pH7.4), 1 mM EDTA and proteaseinhibitors, and are resuspended in the same buffer supplemented with 10%sucrose. Frozen aliquots are kept at −80° C. Competition assays arestarted by mixing 10 nM ³H-ZM214385 (Tocris Cookson) with variousconcentrations of test compounds and 50 μg membrane proteins in TEbuffer (50 mM Tris and 1 mM EDTA) supplemented with 1 Unit/mL adenosinedeaminase. The assays are incubated for 90 minutes, stopped byfiltration using Packard Harvester and washed four times with ice-coldTM buffer (10 mM Tris, 1 mM MgCl2, pH 7.4). Non specific binding isdetermined in the presence of 10 μM ZM214385. The affinities ofcompounds (i.e. Ki values) are calculated using GraphPad software.

Radioligand Binding for Other Adenosine receptors.

Human A₁, A_(2A), A₃ adenosine receptor cDNAs are stably transfectedinto either CHO or HEK-293 cells (referred to as CHO-A1, HEK-A2A,CHO-A3). Membranes are prepared from these cells using the same protocolas described above. Competition assays are started by mixing 0.5 nM³H-CPX (for CHO-A1), 2 nM ³H-ZM214385 (HEK-A2A) or 0.1 nM ¹²⁵I-AB-MECA(CHO-A3) with various concentrations of test compounds and theperspective membranes in TE buffer (50 mM Tris and 1 mM EDTA fo CHO-A1and HEK-A2A) or TEM buffer (50 mM Tris, 1 mM EDTA and 10 mM MgCl₂ forCHO-A3) supplemented with 1 Unit/mL adenosine deaminase. The assays areincubated for 90 minutes, stopped by filtration using Packard Harvesterand washed four times with ice-cold TM buffer (10 mM Tris, 1 mM MgCl2,pH 7.4). Non specific binding is determined in the presence of 1 μM CPX(CHO-A1), 1 μM ZM214385 (HEK-A2A) and 1 μM IB-MECA (CHO-A3). Theaffinities of compounds (i.e. Ki values) are calculated using GraphPadsoftware.

cAMP Measurements.

Monolayer of transfected cells is collected in PBS containing 5 mM EDTA.Cells are washed once with DMEM and resuspended in DMEM containing 1Unit/mL adenosine deaminase at a density of 100,000-500,000 cells/ml.100 μl of the cell suspension is mixed with 25 μl containing variousagonists and/or antagonists and the reaction is kept at 37° C. for 15minutes. At the end of 15 minutes, 125 μl 0.2N HCl is added to stop thereaction. Cells are centrifuged for 10 minutes at 1000 rpm. 100 μl ofthe supernatant is removed and acetylated. The concentrations of cAMP inthe supernatants is measured using the direct cAMP assay from AssayDesign.

A_(2A) and A_(2B) adenosine receptors are coupled to Gs proteins andthus agonists for A_(2A) adenosine receptor (such as CGS21680) or forA_(2B) adenosine receptor (such as NECA) increase the cAMP accumulationswhereas the antagonists to these receptors prevent the increase in cAMPaccumulations-induced by the agonists. A₁ and A₃ adenosine receptors arecoupled to Gi proteins and thus agonists for A₁ adenosine receptor (suchas CPA) or for A₃ adenosine receptor (such as IB-MECA) inhibit theincrease in cAMP accumulations-induced by forskolin. Antagonists to A₁and A₃ receptors prevent the inhibition in cAMP accumulations.

EXAMPLE 15 Comparison of Bioavailability of A_(2B) Adenosine ReceptorProdrugs vs A_(2B) Adenosine Receptor Antagonist

The following studies were conducted in order to compare thepharmacokinetics of the parent A_(2B) adenosine receptor antagonist andits corresponding prodrug. The parent compound chosen was the compoundof formula A in which R¹ is n-propyl, R² is ethyl, R³ is hydrogen, andR⁴ is 3-trifluoromethylphenyl; that is:

where Z is hydrogen (compound 1).

The prodrugs chosen for comparison were as follows:

-   where Z is —CH₂—O—C(O)CH₂CH₂CH₃ (compound ²);-   where Z is —CH₂—O—C(O)CH₃ (compound 3);-   where Z is —CH₂—O—C(O)C(CH₃)₃ (compound ⁴);-   where Z is —CH₂—O—C(O)N(CH₃)CH₂CH₂N(CH₃)₂ (compound 5);-   where Z is —CH₂—O—P(O)(OH)₂ (compound 6)    where Z is

where Z is

where Z is

-   where Z is —CH₂—O—C(O)NHCH₂CH₂N(CH₃)₂ (compound 10);-   where Z is —CH₂—OCH₃ (compound 11) and;-   where Z is methyl (compound 12).

The studies were carried out in Sprague Dawley rats. The test compoundswere administered orally by gavage to groups of three rats using asingle oral dose of the test compound at 2 and 30 mg/kg. All oral doseswere prepared either as a suspension in DMSO/ethanol/PEG300/0.1%N-methyl-D-glucamine or as a suspension in 0.5% methylcellulose inwater. Blood samples were obtained serially from each rat at 0, 5, 15,30 min, and then 1, 1.5, 2, 4, 6, 8, and 24 hrs post-dose.

Determination of Concentrations of Compound 1 and the CorrespondingProdrug in Plasma

Concentrations of compound 1 and/or the corresponding prodrug in ratplasma were determined by a HPLC tandem mass spectrometric (LC/MS/MS)method. Briefly, 0.1 mL of plasma sample was treated with 0.5 mL ofacetonitrile:methanol (9:1, v/v) mixture containing 25 ng of compound 1having deuterated ethyl at the 3-position in place of ethyl. (InternalStandard, I.S.) to precipitate the protein. The mixture was filteredthrough a 96-well filter and filtrate was collected and evaporated todryness on a 96-well plate evaporator. The residue was thenreconstituted with 400 μL 20% methanol and subjected to LC/MS/MSanalysis. Quantification of compound 1 was achieved by mass spectrometryusing Multiple Reaction Monitoring (MRM) mode, monitoring thetransitions at m/z 447.1>159.1 for compound 1 and 452.1>159.1 for I.S.The quantification limit of the assay was 0.38 ng/mL for the analysis ofthe oral dose samples and 10 ng/mL during the analysis of theintravenous dose samples using 0.1 mL of plasma.

Pharmacokinetic Analysis

Non-compartmental pharmacokinetic parameters were determined using acommercial program WinNonLin Professional, Version 4.1 (Pharsight,Mountain View, Calif.). Plasma concentration at below level of detectionwas assumed to be zero for the calculation of means and pharmacokineticparameters.

For oral administration, the maximum concentration (C_(max)) and time toreach C_(max) (T_(max)), AUC₍₀₋₁₎, AUC₍₀₋₈₎ and bioavailability (% F)were determined. Oral bioavailability was determined from the ratio ofdose-adjusted AUC₍₀₋₈₎ of the respective oral dose and the mean AUC₍₀₋₈₎values of the 0.1 and 0.5 mg/kg intravenous doses.

The results are presented below in tabular form. The table providesresults obtained by using a suspension of the test compound in asuspension in DMSO/ethanol/PEG300/0.1% N-methyl-D-glucamine in rats.

Prodrug Compound 1 Mean Dose Mean Cmax Mean Dose Mean Cmax CompoundAdjusted AUC (ng/ml) Adjusted AUC (ng/ml) Compound 1 Not applicable Notapplicable 700 1,900 Compound 2 Not detectable <5 ng/ml 13,790 3,220Compound 3 Not detectable <5 ng/ml 9,993 1,767 Compound 4 Not detectable<5 ng/ml 5,476 1,008 Compound 5 Not detectable <5 ng/ml 2,021 466Compound 6 Not detectable <5 ng/ml 11,800 28,700 Compound 7 51.0 30.01,817 408 Compound 8 Not detectable <5 ng/ml 1,089 241 Compound 9 4.576.39 1,062 160 Compound 10 Not detectable <5 ng/ml 384 36.9 Compound 11533 158 Not detectable <5 ng/ml Compound 12 926 279 Not detectable <5ng/mlWhen administration was carried out in a suspension in 0.5%methylcellulose in water in rats at 30 mg/kg, compound 6 provided a doseadjusted AUC of compound 1 of 11,800 ng/hr/ml, and a Cmax of 28,700ng/ml. Compound 2 provided a dose adjusted AUC of compound 1 of 8,300ng/hr/ml, and a Cmax of 19,200 ng/ml Compound 1 itself provided a doseadjusted AUC of compound 1 of 700 ng/hr/ml, and a Cmax of 1,900 ng/ml.

Results

It can be seen from the results shown above that compounds 2-6 have theideal bioavailability profile of providing much higher plasma levels ofthe parent A_(2B) adenosine receptor antagonist (compound 1) followingoral dosing than is obtained by oral dosing of the parent compound(formula 1) itself Additionally, no trace of the prodrug is seen inplasma. This is in marked contrast to compounds 6-11, which provideplasma levels of the parent A_(2B) adenosine receptor antagonistfollowing oral dosing that are lower than those obtained by oral dosingof the parent compound (formula 1) itself, and, in addition, compounds6,8 and 10-11 are detected in the plasma unmetabolized. Compound 5provided a bioavailability profile that is approximately the same asparent A_(2B) adenosine receptor antagonist (compound 1).

1. A compound of the formula:

wherein: R¹ and R² are independently lower alkyl; R⁴ is optionallysubstituted phenyl; X is hydrogen or methyl; and Y is —C(O)R, in which Ris optionally substituted aryl or optionally substituted heteroaryl; ora pharmaceutically acceptable salt thereof.
 2. The compound of claim 1,wherein R¹ and R² are independently ethyl or n-propyl.
 3. The compoundof claim 1, wherein R⁴ is 3-(trifluoromethyl)phenyl.
 4. The compound ofclaim 3, wherein R¹ is n-propyl and R² is ethyl.
 5. The compound ofclaim 4, wherein X is hydrogen.
 6. (canceled)
 7. (canceled) 8.(canceled)
 9. (canceled)
 10. (canceled)
 11. (canceled)
 12. (canceled)13. A method of treating a disease state chosen from diarrhea and asthmain a mammal, comprising administering to the mammal in need thereof atherapeutically effective dose of a compound of claim
 1. 14. (canceled)15. (Canceled)
 16. The method of claim 13, wherein the disease state isasthma.
 17. (canceled)
 18. The method of claim 13, wherein the diseasestate is diarrhea.
 19. (canceled)
 20. (canceled)
 21. A pharmaceuticalcomposition comprising at least one pharmaceutically acceptableexcipient and a therapeutically effective amount of a compound ofclaim
 1. 22. The compound of claim 5, wherein R is phenyl or pyridyl.23. The compound of claim 22, selected from the group consisting of[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methylbenzoate;[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl3-(trifluoromethyl)benzoate;[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl4-piperazinylbenzoate;[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methyl4-morpholin-4-ylbenzoate and[3-ethyl-2,6-dioxo-1-propyl-8-(1-{[3-(trifluoromethyl)phenyl]methyl}pyrazol-4-yl)-1,3,7-trihydropurin-7-yl]methylpyridine-3-carboxylate.